Estrogen and xenoestrogens upregulate the brain aromatase isoform (P450aromB) and perturb markers of early development in zebrafish (Danio rerio).

Estrogen synthesized in the brain itself by the action of cytochrome P450 aromatase (P450arom) is known to have permanent organizing effects on the developing CNS. In fish, estrogen upregulates the predominant brain isoform (P450aromB), implying that xenoestrogens (XE) could act as neurodevelopmental toxicants by altering P450aromB. To test this hypothesis, zebrafish embryos were exposed to 17beta-estradiol (E(2)), diethylstilbestrol (DES, a potent agonist), and bisphenol A (BPA, a weak agonist). RT-PCR/Southern transfer analysis showed that E(2) (0.01-10 microM) upregulated P450aromB in a dose-response manner. The effect of DES (0.01 microM) was similar to 1 microM E(2) (three- to four-fold higher than control), but BPA was less effective (<threefold increase at 10 microM). mRNA levels of the predominant ovarian isoform (P450aromA) were unchanged by estrogen. Treatment with E(2) (0.1-10 microM) between 2 and 72 hpf had dose-response effects on mortality and hatching and induced a 'curved tail down' phenotype characteristic of mutants with defects of early CNS development. The critical period of estrogen sensitivity for effects on mortality and curved tails was 2-24 hpf, whereas hatching effects were both stage- and duration-dependent. Developmental effects of DES and BPA were similar to E(2) but testosterone, and 5alpha-dihydrotestosterone were ineffective. 17alpha-Estradiol showed a small but significant effect on curved tails. We conclude that P450aromB mRNA is a sensitive marker of XE effect during embryogenesis, but further studies are required to determine whether changes in neural aromatase expression and estrogen biosynthesis have consequences for CNS development.